Preservation of epidermal melanocyte integrity in an ex vivo co-culture skin model with sebocytes.
3D-SeboSkin
SZ95 sebocytes
co-culture techniques
epidermal melanocytes
ex vivo
keratinocytes
skin model
skin organ culture
Journal
Experimental dermatology
ISSN: 1600-0625
Titre abrégé: Exp Dermatol
Pays: Denmark
ID NLM: 9301549
Informations de publication
Date de publication:
07 2023
07 2023
Historique:
revised:
01
02
2023
received:
21
10
2022
accepted:
02
04
2023
medline:
11
7
2023
pubmed:
8
6
2023
entrez:
7
6
2023
Statut:
ppublish
Résumé
A direct contact co-culture of skin explants to SZ95 sebocytes (3D-SeboSkin) has been shown to preserve the integrity of epidermal keratinocytes and dermis. In this study, the properties of epidermal melanocytes were evaluated in the same 3D SeboSkin ex vivo model. Skin explants (n = 6) were maintained in the 3D-SeboSkin model, in direct contact to fibroblasts and alone in serum-free medium (SFM). Histopathological, immunohistochemical, apoptosis and oil red staining evaluations were performed at Days 0 and 6 of incubation. Results revealed preservation and prominent proliferation of basal keratinocytes of the skin explants in addition to preservation of dermal collagen and vasculature at Day 6 in the 3D-SeboSkin culture model and to a lesser extent in co-culture with fibroblasts but not in SFM alone. Melan-A+/Ki67- epidermal melanocytes remained attached to the dermis even at sites of epidermal detachment in the three skin explant models tested. However, the number of epidermal melanocytes was significantly conserved in 3D-SeboSkin cultures in comparison with skin explants in SFM (p < 0.05), whereas no difference was found in comparison with the co-culture with fibroblasts. Few DAPI/TUNEL+ apoptotic melanocytes could mostly be observed in SFM-incubated skin explants. Furthermore, only SZ95 sebocytes in contact to skin explants in 3D-SeboSkin exhibited increased lipogenesis with accumulation of abundant lipid droplets. These results denote that the 3D-SeboSkin model yielded significant preservation of epidermal melanocytes and hence it is optimal for ex vivo studies of abnormalities of skin pigmentation, melanocyte neoplasms and effects of different hormones, cytokines, carcinogens and various therapeutics in a pattern that recapitulates the in vivo environment.
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1063-1071Informations de copyright
© 2023 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Références
Abdel-Naser MB, Abdallah M, de Almeida HL, Jr WU. Human skin cell culture and its impact on Dermatology. 2005 https://www.researchgate.net/publication/234076582_Human_Skin_Cell_Culture_and_its_Impact_on_Dermatology. Accessed December 7, 2022
Coulomb B, Dubertret L. Skin cell culture and wound healing. Wound Repair Regen. 2002;10:109-112.
Abdel-Naser MB, Krasagakis K, Garbe C, Eberle J. Direct effects on proliferation, antigen expression and melanin synthesis of cultured normal human melanocytes in response to UVB and UVA light. Photodermatol Photoimmunol and Photomed. 2003;19:122-127.
Goldstein NB, Koster MI, Hoaglin LG, et al. Narrow band ultraviolet B treatment for human vitiligo is associated with proliferation, migration, and differentiation of melanocyte precursors. J Invest Dermatol. 2015;135:2068-2076.
Abdel-Naser MB, Wollina U, El Okby M, El Sheimy S. PUVA induced lentigines arising in vitiligo: involvement of vitiliginous and normal appearing skin. Clin Exp Dermatol. 2004;29:380-382.
Imokawa G. Autocrine and paracrine regulation of melanocytes in human skin and in pigmentary disorders. Pigment Cell Res. 2004;17:96-110.
Yuan XH, Jin ZH. Paracrine regulation of melanogenesis. Br J Dermatol. 2018;178:632-639.
Nordlund JJ. The melanocyte and the epidermal melanin unit: an expanded concept. Dermatol Clin. 2007;25:271-281.
Upadhyay PR, Ho T, Abdel-Malek ZA. Participation of keratinocyte- and fibroblast-derived factors in melanocyte homeostasis, the response to UV, and pigmentary disorders. Pigment Cell Melanoma Res. 2021;34:762-776.
Kim M, Shibata T, Kwon S, Park TJ, Kang HY. Ultraviolet-irradiated endothelial cells secrete stem cell factor and induce epidermal pigmentation. Sci Rep. 2018;8:4235.
Abdel-Naser MB, Hann SK, Bystryn JC. Oral psoralen with UVA therapy releases circulating growth factor(s) that stimulate cell proliferation. Arch Dermatol. 1997;133:1530-1533.
Abdel-Naser MB, Seltmann H, Altenburg A, Zouboulis CC. Endothelins and α-melanocyte-stimulating hormone are increased in plasma of patients treated with UVA1 and psoralen plus UVA. Photodermatol Photoimmunol Photomed. 2022;38:611-613.
Mazzoleni G, Di Lorenzo D, Steimberg N. Modelling tissues in 3D: the next future of pharmaco-toxicology and food research? Genes Nutr. 2009;4:13-22.
Varani J, Perone P, Spahlinger DM, et al. Human skin in organ culture and human skin cells (keratinocytes and fibroblasts) in monolayer culture for assessment of chemically induced skin damage. Toxicol Pathol. 2007;35:693-701.
Lebonvallet N, Jeanmaire C, Danoux L, Sibille P, Pauly G, Misery L. The evolution and use of skin explants: potential and limitations for dermatological research. Eur J Dermatol. 2010;20:671-684.
Jang YH, Kim SL, Lee JS, et al. Possible existence of melanocytes or melanoblasts in human sebaceous glands. Ann Dermatol. 2014;26:469-473.
Abdel-Naser MB, Seltmann H, Zouboulis CC. SZ95 sebocytes induce epidermal melanocyte dendricity and proliferation in vitro. Exp Dermatol. 2012;21:393-395.
Flori E, Mastrofrancesco A, Mosca S, et al. Sebocytes contribute to melasma onset. iScience. 2022;25:103871.
Casalou C, Moreiras H, Mayatra JM, Fabre A, Tobin DJ. Loss of ‘epidermal melanin unit’ integrity in human skin during melanoma-genesis. Front Oncol. 2022;12:878336.
Nikolakis G, Seltmann H, Hossini AM, Makrantonaki E, Knolle J, Zouboulis CC. Ex vivo human skin and SZ95 sebocytes exhibit a homoeostatic interaction in a novel co-culture contact model. Exp Dermatol. 2015;24:497-502.
Zouboulis CC, Seltmann H, Neitzel H, Orfanos CE. Establishment and characterization of an immortalized human sebaceous gland cell line (SZ95). J Invest Dermatol. 1999;113:1011-1020.
Hou X, Hossini AM, Nikolakis G, Balthasar O, Kurtz A, Zouboulis CC. 3D-SeboSkin model for human ex vivo studies of hidradenitis suppurativa/acne inversa. Dermatology. 2022;238:236-243.
Zouboulis CC, Hou X, von Waldthausen H, Zouboulis KC, Hossini AM. HS 3D-SeboSkin model enables the preclinical exploration of therapeutic candidates for hidradenitis suppurativa/acne inversa. Pharmaceutics. 2023;15:619.
Prieto VG, Shea CR. Immunohistochemistry of melanocytic proliferations. Arch Pathol Lab Med. 2011;135:853-859.
Abdallah MA, Abdel-Naser MB, Moussa MH, Assaf C, Orfanos CE. Sequential immuno-histochemical study of depigmenting and repigmenting minigrafts in vitiligo. Eur J Dermatol. 2003;13:548-552.
Abdel-Naser MB. Mitogen requirements of normal epidermal human melanocytes in a serum and tumor promoter free medium. Eur J Dermatol. 2003;13:29-33.
Adigun R, Basit H, Murray J. Necrosis. Cell (Liquefactive, Coagulative, Caseous, Fat, Fibrinoid, and Gangrenous). [Updated 2019 Jun 30]. StatPearls. StatPearls Publishing; 2020 https://www.ncbi.nlm.nih.gov/books/NBK430935/
Kumar V, Abbas AK, Aster J. Robbins and Cotran Pathologic Basis of Disease. 10th ed. Elsevier Saunders; 2018:36.
Hirobe T. How are proliferation and differentiation of melanocytes regulated? Pigment Cell Melanoma Res. 2011;24:462-478.
Abdel-Naser MB, Liakou AI, Elewa R, Hippe S, Knolle J, Zouboulis CC. Increased activity and number of epidermal melanocytes in lesional psoriatic skin. Dermatology. 2016;232:425-430.
Cario-André M, Pain C, Gauthier Y, Casoli V, Taieb A. In vivo and in vitro evidence of dermal fibroblasts influence on human epidermal pigmentation. Pigment Cell Res. 2006;19:434-442.
Gross-Amat O, Guillen M, Salmon D, Nataf S, Auxenfans C. Characterization of a topically testable model of burn injury on human skin explants. Int J Mol Sci. 2020;21:6956.
von Müller C, Bulman F, Wagner L, et al. Active neutrophil responses counteract Candida albicans burn wound infection of ex vivo human skin explants. Sci Rep. 2020;10:21818.
Abdel-Naser MB. Differential effects on melanocyte growth and melanization of low vs high Ca++ keratinocyte conditioned medium. Brit J Dermatol. 1999;140:50-55.
Wang Y, Viennet C, Robin S, Berthon JY, He L, Humbert P. Precise role of dermal fibroblasts on melanocyte pigmentation. J Dermatol Sci. 2017;88:159-166.
Abadie S, Bedos P, Rouquette J. A human skin model to evaluate the protective effect of compounds against UVA damage. Int J Cosmet Sci. 2019;41:594-603.
Sridharan A, Shi M, Leo VI, et al. The polyamine putrescine promotes human epidermal melanogenesis. J Invest Dermatol. 2020;140:2032-2040.e1.
Wolf Horrell E, D'Orazio J. UV-independent induction of beta defensin 3 in neonatal human skin explants. F1000Res. 2014;3:288.
Eberlin S, Silva MSD, Facchini G, et al. The ex vivo skin model as an alternative tool for the efficacy and safety evaluation of topical products. Altern Lab Anim. 2020;48:10-22.
van den Boorn JG, Konijnenberg D, Dellemijn TA, et al. Autoimmune destruction of skin melanocytes by perilesional T cells from vitiligo patients. J Invest Dermatol. 2009;129:2220-2232.